The use of herpes simplex virus-1 vectors in nociceptive biology

摘要: Public Health Relevance: The United States has 80 million employees with chronic pain resulting in annual losses of 61.2 billion dollars due to pain-related productive time lost. In addition, depression and inactivity reduce the quality life. development effective analgesics is therefore important from a public health perspective. this dissertation, natural properties herpes simplex virus (HSV-1) vectors are exploited (i) develop an HSV-1 vector-based selection system that can potentially identify or chemical inhibitors (ii) test vector-expressed dominant negative PKCe (DNP) as strategy treat pain. vanilloid/capsaicin receptor (TRPV1) pro-nociceptive calcium ion channel upregulated This occurs partly protein kinase C epsilon (PKCe)−mediated phosphorylation. An vector expressing TRPV1 (vTT) was engineered vTT-expressed functionality confirmed.Treatment vTT-infected cells capsaicin resiniferatoxin caused concentration-dependent Ca+2 influx, leading cell-death dramatic reduction infectious particle yield. antagonists, ruthenium red SB-366791 reversed agonist-induced rescued vTT growth, providing basis for selection. Selection antagonists modeled using mixed infection vHG (capsaicin resistant control vector) passage presence capsaicin. These experiments demonstrated single readily isolated population 10^5 particles. approach be used gene libraries offers advantages platform assay applicable other channels adaptability high throughput formats. Dominant into create vector, vHDNP. Following functional confirmation vHDNP U2OS, Vero neurons, cobalt uptake showed sensitive vHDNP-transduced neurons. Electrophysiology confirmed also knockdown TRPV1-PKCe coupling nociceptive In-vivo studies noxious heat-induced nocisponsive behavior vHDNP-inoculated rats subtle inhibition withdrawal responses when compared controls. conclusion, expressed viable specifically inhibit function order